From screening to process optimization: AMBR technology to speed up microbial fermentation processes

Session proposals: · Therapeutic Proteins Vaccines The development of biopharmaceuticals or biotechnological products derived from microbial fermentation is a financially risky endeavor and time consuming process, requiring technical upstream solutions which reduce timelines, increase efficiency, and raise likelihood of success. We have identified in particular the early steps of strain and process development offering best prospects to speed up the entire process significantly by using a reliable screening system. Based on the well-proven ambr® principle we designed with ambr 15 fermentation system to accelerate early stage development of microbial fermentation products. The multi-fermentation unit mimics larger scale bioreactor processes, and is suitable for screening clones, strains or growth conditions. In case studies with industrial partners using E. coli and P. pastoris, consistent and efficient control of fermentations across a variety culture conditions (e.g. feed, temperature, duration, pH) could be demonstrated. In the succeeding step of process development ambr 250 has been widely applied to speed up the 2nd critical phase of the microbial upstream process development. The larger working volume and the range of features, which this multi-parallel system offers, are superior to common benchtop fermenters. Optical density supervision, off gas analysis, fed-batch processing and advanced control capabilities allow process development for most commercial-scale upstream fermentation processes. In addition to this impressive range of features ambr250 has proven its ability to reliably increase the efficiency of fermentation process development many times through its rapid setup and cleanup, advanced control software, and automation

Establishment of centromere identity is dependent on nuclear spatial organization

The establishment of centromere-specific CENP-A chromatin is influenced by epigenetic and genetic processes. Central domain sequences from fission yeast centromeres are preferred substrates for CENP-A(Cnp1) incorporation, but their use is context dependent, requiring adjacent heterochromatin. CENP-A(Cnp1) overexpression bypasses heterochromatin dependency, suggesting that heterochromatin ensures exposure to conditions or locations permissive for CENP-A(Cnp1) assembly. Centromeres cluster around spindle-pole bodies (SPBs). We show that heterochromatin-bearing minichromosomes localize close to SPBs, consistent with this location promoting CENP-A(Cnp1) incorporation. We demonstrate that heterochromatin-independent de novo CENP-A(Cnp1) chromatin assembly occurs when central domain DNA is placed near, but not far from, endogenous centromeres or neocentromeres. Moreover, direct tethering of central domain DNA at SPBs permits CENP-A(Cnp1) assembly, suggesting that the nuclear compartment surrounding SPBs is permissive for CENP-A(Cnp1) incorporation because target sequences are exposed to high levels of CENP-A(Cnp1) and associated assembly factors. Thus, nuclear spatial organization is a key epigenetic factor that influences centromere identity

Presentation of SLE in UK primary care using the Clinical Practice Research Datalink

OBJECTIVES: To describe the presenting symptoms of SLE in primary care using the Clinical Practice Research Database (CPRD) and to calculate the time from symptom presentation to SLE diagnosis. METHODS: Incident cases of SLE were identified from the CPRD between 2000 and 2012. Presenting symptoms were identified from the medical records of cases in the 5 years before diagnosis and grouped using the British Isles Lupus Activity Group (BILAG) symptom domains. The time from the accumulation of one, two and three BILAG domains to SLE diagnosis was investigated, stratified by age at diagnosis (<30, 30–49 and ≥50 years). RESULTS: We identified 1426 incident cases (170 males and 1256 females) of SLE. The most frequently recorded symptoms and signs prior to diagnosis were musculoskeletal, mucocutaneous and neurological. The median time from first musculoskeletal symptom to SLE diagnosis was 26.4 months (IQR 9.3–43.6). There was a significant difference in the time to diagnosis (log rank p<0.01) when stratified by age and disease severity at baseline, with younger patients <30 years and those with severe disease having the shortest times and patients aged ≥50 years and those with mild disease having the longest (6.4 years (IQR 5.8–6.8)). CONCLUSIONS: The time from symptom onset to SLE diagnosis is long, especially in older patients. SLE should be considered in patients presenting with flaring or chronic musculoskeletal, mucocutaneous and neurological symptoms